This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This projects focuses on the development of novel grid substrate and cryo-EM specimen preparation technologies. The specific aims of the project are: 1.) Explore and evaluate new designs and geometries for holey carbon foils in order to optimize the efficiency of digital image acquisition and the quality of the images acquired with the goal of improving stability, conductivity and specimen flatness. 2.) Systematically investigate factors that influence the production of large areas of thin ice, as well as the way particles orient and are distributed on the holey grids. 3.) Explore and evaluate novel specimen supports designed to address some of the problems associated with carbon support films. The aim is to provide a new substrate with improved mechanical stability, electrical conductivity and flatness.